التفاصيل البيبلوغرافية
| العنوان: |
In vitro generation of megakaryocytes from engineered mouse embryonic stem cells. |
| المؤلفون: |
Lewis MR; Department of Biomedical Engineering, University of Utah, Salt Lake City, UT 84112, USA., Deans TL; Department of Biomedical Engineering, University of Utah, Salt Lake City, UT 84112, USA. |
| المصدر: |
BioRxiv : the preprint server for biology [bioRxiv] 2023 Mar 01. Date of Electronic Publication: 2023 Mar 01. |
| نوع المنشور: |
Preprint |
| اللغة: |
English |
| بيانات الدورية: |
Country of Publication: United States NLM ID: 101680187 Publication Model: Electronic Cited Medium: Internet NLM ISO Abbreviation: bioRxiv Subsets: PubMed not MEDLINE |
| مستخلص: |
The in vitro differentiation of pluripotent stem cells into desired lineages enables mechanistic studies of cell transitions into more mature states that can provide insights into the design principles governing cell fate control. We are interested in reprogramming pluripotent stem cells with synthetic gene circuits to drive mouse embryonic stem cells (mESCs) down the hematopoietic lineage for the production of megakaryocytes, the progenitor cells for platelets. Here, we describe the methodology for growing and differentiating mESCs, in addition to inserting a transgene to observe its expression throughout differentiation. This entails four key methods: (1) growing and preparing mouse embryonic fibroblasts for supporting mESC growth and expansion, (2) growing and preparing OP9 feeder cells to support the differentiation of mESCs, (3) the differentiation of mESCs into megakaryocytes, and (4) utilizing an integrase mediated docking site to insert transgenes for their stable integration and expression throughout differentiation. Altogether, this approach demonstrates a streamline differentiation protocol that emphasizes the reprogramming potential of mESCs that can be used for future mechanistic and therapeutic studies of controlling cell fate outcomes. |
| التعليقات: |
Update in: Methods Mol Biol. 2024;2774:279-301. (PMID: 38441772) |
| معلومات مُعتمدة: |
DP2 CA250006 United States CA NCI NIH HHS; P30 CA042014 United States CA NCI NIH HHS; S10 OD026959 United States OD NIH HHS |
| تواريخ الأحداث: |
Date Created: 20230313 Latest Revision: 20240318 |
| رمز التحديث: |
20240318 |
| مُعرف محوري في PubMed: |
PMC10002726 |
| DOI: |
10.1101/2023.03.01.530476 |
| PMID: |
36909620 |
| قاعدة البيانات: |
MEDLINE |
